goat anti human type iii collagen  (Bio-Rad)

Journal: Journal of refractive surgery (Thorofare, N.J. : 1995)

Article Title: Regeneration of defective epithelial basement membrane and restoration of corneal transparency

doi: 10.3928/1081597X-20170126-02

Figure Lengend Snippet: Rabbit corneas that are unwounded, and at 2 weeks and 1 month after -9 diopters (D) photorefractive keratectomy (PRK). (A) Control corneas were clear without any haze (mag 25X). (B) Mild haze restricted to the area of excimer laser ablation was noted in all corneas at two weeks after the ablation (mag 25X). (C) Dense subepithelial haze was observed at one month after PRK (mag 25X). (D) Control corneas had no alpha-smooth muscle actin (α-SMA)+ cells (mag 200X). (E) Corneas had a few α-SMA+ cells (arrowheads) in the excimer laser ablated zone at two weeks after -9D PRK (mag 200X). (F) α-SMA+ cells (arrowheads) were present at high density in the subepithelial excimer laser-ablated zone in all corneas after 1 month (mag 200X). (G) Transmission electron microscopy (TEM) of control corneas showed normal epithelial basement membrane (EBM) lamina densa (arrows) and lamina lucida, and normal organized extracellular matrix (mag 23,000X). (H) At two weeks after -9D PRK, there was no detectible lamina lucida and lamina densa where EBM is normally noted (x) within the excimer laser ablated zone and disorganized extracellular matrix was prominent in the subepithelial stroma (s) (mag 23,000X). (I) At one month after -9D PRK, no lamina lucida or lamina densa were detected within the excimer laser ablated zone where EBM would be detected (x) and there were layers of cells (m) with large amounts of rough endoplasmic reticulum that correspond to the α-SMA+ cells (myofibroblasts) in Fig. 1F embedded in disorganized extracellular matrix (d) (mag 23,000X). (J) Control corneas had no detectible collagen type III (COL3) (mag 400X). (K) At two weeks after -9D PRK, corneas had COL3 (arrows) in the subepithelial stroma (mag 400X). (L) At one month after -9D PRK, all corneas had greater COL3 (arrows) in the subepithelial stroma (mag 400X). Blue is DAPI staining of cell nuclei; (e) epithelium; (s) stroma.

Article Snippet: COL3 was detected using a purified goat polyclonal antibody anti-human Collagen α1 Type III (sc-8781, Santa Cruz Biotechnology, Inc. Dallas, TX) that also recognized rabbit antigen.

Techniques: Control, Transmission Assay, Electron Microscopy, Membrane, Staining

Journal: Journal of refractive surgery (Thorofare, N.J. : 1995)

Article Title: Regeneration of defective epithelial basement membrane and restoration of corneal transparency

doi: 10.3928/1081597X-20170126-02

Figure Lengend Snippet: Rabbit corneas at one, two and three months after non-perforating linear incisions. (A) At one month after the surgery, the two incisions (arrowheads) were visible as dense linear opacities in all nine corneas that reached this time point (mag 25X). (B) At two months after surgery, the incisions (arrowheads) were less opaque in all six corneas that reached this time point (mag 25X). (C) At three months after surgery, the incisions (arrowheads) were faint compared to the two-month time point in all three corneas that reached this time point (mag 25X). (D), (E), and (F) At one, two or three months after surgery, respectively, there were no alpha-smooth muscle actin (α-SMA)+ cells anywhere in the cornea, including around the epithelial plugs (arrows), in three corneas at each time point (mag 200X). (G), (H), and (I) At one, two and three months, respectively, after surgery the EBM (arrows), including lamina lucida and lamina densa, were fully regenerated and surrounded the epithelial plugs in all three corneas at each time point (mag 23,000X). (J) At 1 month after surgery, both incision in each of the three corneas studied had collagen type III (COL3, arrows) deposited in the subepithelial stroma beneath the epithelial plugs (mag 200X). At two months (K) and three months (L) after surgery, more COL3 deposition (arrows), compared to the one month time point, was noted in the subepithelial stroma surrounding the epithelial plugs in each of the three corneas at each time points. Blue is DAPI staining of cell nuclei; (e) epithelium; (s) stroma; (*) indicates artifact separation of the epithelium from the stroma that occurs during cutting of the section.

Article Snippet: COL3 was detected using a purified goat polyclonal antibody anti-human Collagen α1 Type III (sc-8781, Santa Cruz Biotechnology, Inc. Dallas, TX) that also recognized rabbit antigen.

Techniques: Staining